A cell cycle alteration precedes apoptosis of granule cell precursors in the weaver mouse cerebellum

Antonio Migheli, Roberto Piva, Stefania Casolino, Cristiana Atzori, Stephen Dlouhy, Bernardino Ghetti

Research output: Contribution to journalArticle

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Abstract

A missense mutation in the gene coding for the G-protein-activated inwardly rectifying potassium (GIRK) channel, GIRK2, is responsible for apoptosis in the external germinal layer (EGL) of the cerebellum and a nonapoptotic death of midbrain dopaminergic neurons in the weaver (wv) mouse. Failure of axonogenesis and migration are considered to be the primary consequences of GIRK2 channel malfunction in the cerebellum. We investigated whether a disruption of the cell cycle precedes the failure of migration and axonogenesis and leads to massive apoptosis. To this end, immunohistochemistry and immunoblotting for PCNA, Cdk4, cyclin D, cyclin A, and the Cdk inhibitor p27/kip1, as well as in situ end-labeling for apoptotic DNA fragmentation, were applied to cerebella of P7-P21+/+, wv/+, and wv/wv mice. In +/+ and wv/+ mice, the expression of cell cycle proteins was limited to the outer, premigratory zone of the EGL. Antibodies to p27, a marker of cell differentiation, gave a reverse staining pattern. Due to migration delay, patches of p27-positive cells persisted in the outer EGL in P21 wv/+ mice. On the contrary, marked cell cycle up-regulation and absence of p27 occurred throughout the EGL at all ages in wv/wv mice, indicating an inability to switch off the cell cycle. Mitotic index evaluation showed that cell cycle activation was unrelated to proliferative events. Cell cycle proteins were not expressed in the substantia nigra, suggesting that nonapoptotic death of mature dopaminergic neurons is not preceded by abortive cell cycle reentry. Our data show that abnormalities of the cell cycle in wv/wv cerebellum represent a major and early consequence of GIRK2 channel malfunction and may strongly influence the susceptibility of EGL cells to apoptosis. These observations may help in understanding the pathogenesis of human neurological channelopathies.

Original languageEnglish
Pages (from-to)365-373
Number of pages9
JournalAmerican Journal of Pathology
Volume155
Issue number2
StatePublished - Aug 1999

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Neurologic Mutant Mice
Cerebellum
Cell Cycle
Apoptosis
Cell Cycle Proteins
Dopaminergic Neurons
G Protein-Coupled Inwardly-Rectifying Potassium Channels
Channelopathies
Cyclin D
Inwardly Rectifying Potassium Channel
Cyclin A
Mitotic Index
Proliferating Cell Nuclear Antigen
DNA Fragmentation
Substantia Nigra
Missense Mutation
Mesencephalon
Immunoblotting
Cell Differentiation
Up-Regulation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

A cell cycle alteration precedes apoptosis of granule cell precursors in the weaver mouse cerebellum. / Migheli, Antonio; Piva, Roberto; Casolino, Stefania; Atzori, Cristiana; Dlouhy, Stephen; Ghetti, Bernardino.

In: American Journal of Pathology, Vol. 155, No. 2, 08.1999, p. 365-373.

Research output: Contribution to journalArticle

Migheli, Antonio ; Piva, Roberto ; Casolino, Stefania ; Atzori, Cristiana ; Dlouhy, Stephen ; Ghetti, Bernardino. / A cell cycle alteration precedes apoptosis of granule cell precursors in the weaver mouse cerebellum. In: American Journal of Pathology. 1999 ; Vol. 155, No. 2. pp. 365-373.
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